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INTESTINAL Publication-derived

Human Intestinal Organoids Model Cryptosporidium Infection

Source Heo et al. · Cincinnati Children's Hospital Medical Center, Cincinnati, USA · 10.1038/s41564-018-0177-8

👤 Heo I, Dutta D, Schaefer DA, Iakobachvili N, Artegiani B, Sachs N, Boonekamp KE, Bowber G, Huber AR, Riggs MW, Clevers H ⏱ 72 days 📋 7 phases 🧫 iPSC-Derived (Human iPSC-Derived Intestinal Organoids)

Abstract

Achieved the first complete Cryptosporidium parvum life cycle in human intestinal organoids through microinjection of oocysts into the organoid lumen. This breakthrough enabled long-term culture of a previously unculturable parasite and established a platform for anti-parasitic drug screening.

Cell source
iPSC-Derived (Human iPSC-Derived Intestinal Organoids)
Application
Parasitic Infection Modeling

Protocol overview

19 steps across 7 phases

Organoid Culture Preparation Days 0–7
  1. 1 SI Organoid Culture Setup
  2. 2 SI Organoid Differentiation
  3. 3 Lung Organoid Culture Setup
  4. 4 Prepare Organoids for Microinjection
Cryptosporidium parvum Preparation Day 0
  1. 1 Oocyst Preparation for Microinjection
  2. 2 Sporozoite Preparation for Microinjection
Microinjection and Infection Day 0
  1. 1 Prepare Microinjection Apparatus
  2. 2 Perform Microinjection
  3. 3 Culture Infected Organoids
Long-Term Culture (Optional) Days 0–28
  1. 1 Switching Differentiation Protocol for Parasite Maintenance
Parasite Detection and Quantification Days 0–6
  1. 1 RNA Extraction and qRT-PCR
  2. 2 Immunofluorescence Assay (IFA)
  3. 3 Electron Microscopy—Scanning Electron Microscopy (SEM)
  4. 4 Electron Microscopy—Transmission Electron Microscopy (TEM)
Oocyst Isolation and Viability Testing Days 4–6
  1. 1 Isolate Oocysts from Organoids
  2. 2 Mouse Infectivity Assay (Optional but Recommended)
RNA Sequencing and Transcriptome Analysis Days 0–72
  1. 1 Sample Preparation for RNA Sequencing
  2. 2 Library Preparation and Sequencing
  3. 3 Transcriptome Data Analysis

Full SOP

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Attribution

This SOP was authored by Organthis based on the published method in Heo et al.. The originating laboratory holds no rights in this SOP and has not endorsed it unless marked Verified.

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